METHODS OF USING CHIMERIC ANTIGEN RECEPTORS TARGETING B-CELL MATURATION ANTIGEN AND USES THEREOF
The presently disclosed subject matter provides for methods and compositions for treating multiple myeloma. It relates to chimeric antigen receptors (CARs) that specifically target B cell maturation antigen (BCMA), and immunoresponsive cells comprising such CARs. The presently disclosed BCMA-specific CARs have enhanced immune-activating properties, including anti-tumor activity.
1 . A chimeric antigen receptor (CAR), comprising an extracellular antigen-binding domain that specifically binds to B cell maturation antigen (BCMA), a transmembrane domain and an intracellular domain, wherein the extracellular antigen-binding domain comprises a heavy chain variable region and a light chain variable region, wherein:
(a) the heavy chain variable region comprises an amino acid sequence that is at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to an amino acid sequence selected from the group consisting of: SEQ ID NOS: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, and 65; and/or the light chain variable region comprises an amino acid sequence that is at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, and 66; and/or
(b) the heavy chain variable region comprises a CDR1, a CDR2, and a CDR3 of a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, or 65, and/or the light chain variable region comprises a CDR1, a CDR2, and a CDR3 of a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, or 66; and/or
(c) the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 91, 97, 103, 109, 115, 121, 127, 133, 139, 145, 151, 157, 163, 169, 175, 181, and 187, and conservative modifications thereof; and/or the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 94, 100, 106, 112, 118, 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, and 190, and conservative modifications thereof.
2 . The CAR of claim 1 , wherein the extracellular antigen-binding domain of the CAR binds to B cell maturation antigen (BCMA) with a binding affinity (K D ) of from about 1×10 −9 M to about 3 ×10 −6 M.
3 . The CAR of claim 1 , wherein the extracellular antigen-binding domain of the CAR binds to B cell maturation antigen (BCMA) with a binding affinity (K D ) of from about 1×10 −9 M to about 1 ×10 −8 M.
4 . The CAR of claim 1 , wherein
(a) the heavy chain variable region comprises an amino acid sequence that is at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to an amino acid sequence selected from the group consisting of: SEQ ID NOS: 21, 53, 57, 61, and 65; and/or
(b) the light chain variable region comprises an amino acid sequence that is at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOS: 22, 54, 58, 62, and 66.
5 . The CAR of claim 1 , wherein
(a) the heavy chain variable region comprises the CDR1, CDR2, and CDR3 of a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21, 53, 57, 61, or 65; and
(b) the light chain variable region comprises the CDR1, CDR2, and CDR3 of a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 22, 54, 58, 62, or 66.
6 . The CAR of claim 1 , wherein
(a) the heavy chain variable region comprises an amino acid sequence selected from the group consisting of: SEQ ID NOS: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, and 65 and conservative modifications thereof; and/or
(b) the light chain variable region comprises an amino acid sequence selected from the group consisting of: SEQ ID NOS: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, and 66 and conservative modifications thereof.
7 . The CAR of claim 6 , wherein
(a) the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 21, SEQ ID NO: 53, SEQ ID NO: 57, SEQ ID NO: 61, or SEQ ID NO: 65; and/or
(b) the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 22, SEQ ID NO: 54, SEQ ID NO: 58, SEQ ID NO: 62 or SEQ ID NO: 6.
8 . The CAR of claim 6 , wherein the extracellular antigen-binding domain comprises:
(a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 1, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 2;
(b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 5, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 6;
(c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 9, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 10;
(d) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 13, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 14;
(e) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 17, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 18;
(f) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 22;
(g) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 25, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 26;
(h) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 29, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 30;
(i) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 33, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 34;
(j) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 37, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 38;
(k) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 41, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 42;
(1) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 45, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 46;
(m) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 49, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 50;
(n) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 53, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 54;
(o) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 57, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 58;
(p) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 62; or
(q) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 65, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 66.
9 . The CAR of claim 8 , wherein the extracellular antigen-binding domain comprises:
(a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 22;
(b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 53; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 54;
(c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 57; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 58;
(d) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 62; or
(e) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 65; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 66.
10 . The CAR of claim 1 , wherein the extracellular antigen-binding domain comprises a linker between the heavy chain variable region and the light chain variable region.
11 . The CAR of claim 1 , wherein (a) the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 91, 97, 103, 109, 115, 121, 127, 133, 139, 145, 151, 157, 163, 169, 175, 181, and 187, and conservative modifications thereof; and (b) the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 94, 100, 106, 112, 118, 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, and 190, and conservative modifications thereof.
12 . The CAR of claim 11 , wherein (a) the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 90, 96, 102, 108, 114, 120, 126, 132, 138, 144, 150, 156, 162, 168, 174, 180, and 186, and conservative modifications thereof; and (b) the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 93, 99, 105, 111, 117, 123, 129, 135, 141, 147, 153, 159, 165, 171, 177, 183, and 189, and conservative modifications thereof.
13 . The CAR of claim 11 , wherein (a) the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 89, 95, 101, 107, 113, 119, 125, 131, 137, 143, 149, 155, 161, 167, 173, 179, and 185, and conservative modifications thereof; and (b) the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 92, 98, 104, 110, 116, 122, 128, 134, 140, 146, 152, 158, 164, 170, 176, 182, and 188, and conservative modifications thereof.
14 . The CAR of claim 1 , wherein the heavy chain variable region comprises:
(a) the amino acid sequence set forth in SEQ ID NO: 89 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 90 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 91 or a conservative modification thereof;
(b) the amino acid sequence set forth in SEQ ID NO: 95 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 96 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 97 or a conservative modification thereof;
(c) the amino acid sequence set forth in SEQ ID NO: 101 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 102 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 103 or a conservative modification thereof;
(d) the amino acid sequence set forth in SEQ ID NO: 107 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 108 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 109 or a conservative modification thereof;
(e) the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof;
(f) the amino acid sequence set forth in SEQ ID NO: 119 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 120 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 121 or a conservative modification thereof;
(g) the amino acid sequence set forth in SEQ ID NO: 125 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 126 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 127 or a conservative modification thereof;
(h) the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof;
(i) the amino acid sequence set forth in SEQ ID NO: 137 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 138 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 139 or a conservative modification thereof;
(j) the amino acid sequence set forth in SEQ ID NO: 143 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 144 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 145 or a conservative modification thereof;
(k) the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof;
(1) the amino acid sequence set forth in SEQ ID NO: 155 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 156 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 157 or a conservative modification thereof;
(m) the amino acid sequence set forth in SEQ ID NO: 161 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 162 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 163 or a conservative modification thereof;
(n) the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof;
(o) the amino acid sequence set forth in SEQ ID NO: 173 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 174 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 175 or a conservative modification thereof;
(p) the amino acid sequence set forth in SEQ ID NO: 179 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 180 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 181 or a conservative modification thereof; or
(q) the amino acid sequence set forth in SEQ ID NO: 185 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 186 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 187 or a conservative modification thereof.
15 . The CAR of claim 14 , wherein the heavy chain variable region comprises:
(a) the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof;
(b) the amino acid sequence set forth in SEQ ID NO: 173 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 174 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 175 or a conservative modification thereof;
(c) the amino acid sequence set forth in SEQ ID NO: 185 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 186 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 187 or a conservative modification thereof;
(d) the amino acid sequence set forth in SEQ ID NO: 119 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 120 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 121 or a conservative modification thereof; or
(e) the amino acid sequence set forth in SEQ ID NO: 179 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 180 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 181 or a conservative modification thereof.
16 . The CAR of claim 1 , wherein the light chain variable region comprises:
(a) the amino acid sequence set forth in SEQ ID NOs: 92 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 93 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 94 or a conservative modification thereof;
(b) the amino acid sequence set forth in SEQ ID NO: 98 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 99 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 100 or a conservative modification thereof;
(c) the amino acid sequence set forth in SEQ ID NO: 104 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 105 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 106 or a conservative modification thereof;
(d) the amino acid sequence set forth in SEQ ID NO: 110 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 111 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 112 or a conservative modification thereof;
(e) the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof;
(f) the amino acid sequence set forth in SEQ ID NO: 122 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 123 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 124 or a conservative modification thereof;
(g) the amino acid sequence set forth in SEQ ID NO: 128 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 129 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 130 or a conservative modification thereof;
(h) the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof;
(i) the amino acid sequence set forth in SEQ ID NO: 140 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 141 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 142 or a conservative modification thereof;
(j) the amino acid sequence set forth in SEQ ID NO: 146 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 147 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 148 or a conservative modification thereof;
(k) the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 153 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof;
(1) the amino acid sequence set forth in SEQ ID NO: 158 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 159 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 160 or a conservative modification thereof;
(m) the amino acid sequence set forth in SEQ ID NO: 164 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 165 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 166 or a conservative modification thereof;
(n) the amino acid sequence set forth in SEQ ID NO: 170 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 171 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 172 or a conservative modification thereof;
(o) the amino acid sequence set forth in SEQ ID NO: 176 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 177 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 178 or a conservative modification thereof;
(p) the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 183 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 184 or a conservative modification thereof; or
(q) the amino acid sequence set forth in SEQ ID NO: 188 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 189 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 190 or a conservative modification thereof.
17 . The CAR of claim 16 , wherein the light chain variable region comprises:
(a) the amino acid sequence set forth in SEQ ID NO: 170 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 171 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 172 or a conservative modification thereof;
(b) the amino acid sequence set forth in SEQ ID NO: 176 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 177 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 178 or a conservative modification thereof;
(c) the amino acid sequence set forth in SEQ ID NO: 188 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 189 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 190 or a conservative modification thereof;
(d) the amino acid sequence set forth in SEQ ID NO: 122 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 123 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 124 or a conservative modification thereof; or
(e) the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof, the amino acid sequence set forth in SEQ ID NO: 183 or a conservative modification thereof, and the amino acid sequence set forth in SEQ ID NO: 184 or a conservative modification thereof.
18 . The CAR of claim 1 , wherein the extracellular antigen-binding domain comprises:
(a) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 89, 90, and 91; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 92, 93, and 94;
(b) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 95, 96, and 97; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 98, 99, and 100;
(c) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 101, 102, and 103; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 104, 105, and 106;
(d) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 107, 108, and 109; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 110, 111, and 112;
(e) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 113, 114, and 115; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 116, 117, and 118;
(f) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 119, 120, and 121; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 122, 123, and 124;
(g) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 125, 126, and 127; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 128, 129, and 130;
(h) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 131, 132, and 133; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 134, 135, and 136;
(i) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 137, 138, and 139; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 140, 141, and 142;
(j) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 143, 144, and 145; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 146, 147, and 148;
(k) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 149, 150, and 151; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 152, 153, and 154;
(1) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 155, 156, and 157; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 158, 159, and 160;
(m) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 161, 162, and 163; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 164, 165, and 166;
(n) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 167, 168, and 169; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 170, 171, and 172;
(o) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 173, 174, and 175; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 176, 177, and 178;
(p) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 179, 180, and 181; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 182, 183, and 184; or
(q) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 185, 186, and 187; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 188, 189, and 190.
19 . The CAR of claim 18 , wherein the extracellular antigen-binding domain comprises:
(a) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 167, 168, and 169; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 170, 171, and 172;
(b) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 173, 174, and 175; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 176, 177, and 178;
(c) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 185, 186, and 187; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 188, 189, and 190;
(d) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 119, 120, and 121; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 122, 123, and 124; or
(e) a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 179, 180, and 181; and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 182, 183, and 184.
20 . The CAR of claim 1 , wherein
a) the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 21 and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 22;
b) the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 53 and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 54;
c) the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 57 and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 58;
d) the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 61 and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 62; or
e) the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 65 and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 66.
21 . The CAR of claim 1 , wherein the BCMA comprises the amino acid sequence set forth in SEQ ID NO: 71.
22 . The CAR of claim 1 , wherein the extracellular antigen-binding domain binds to an epitope region comprising residues within amino acids 14-22 of SEQ ID NO: 71.
23 . The CAR of claim 1 , wherein the extracellular antigen-binding domain comprises a signal peptide that is covalently joined to the N-terminus of the extracellular antigen-binding domain.
24 . The CAR of claim 1 , wherein the transmembrane domain comprises a CD8 polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a CTLA-4 polypeptide, a PD-1 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide, a synthetic peptide (not based on a protein associated with the immune response), or a combination thereof.
25 . The CAR of claim 24 , wherein the transmembrane domain comprises a CD8 polypeptide or a CD28 polypeptide.
26 . The CAR of claim 1 , wherein the intracellular domain comprises a CD3ζ polypeptide.
27 . The CAR of claim 26 , wherein the intracellular domain comprises at least one signaling region.
28 . The CAR of claim 27 , wherein the at least one signaling region comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, a PD-1 polypeptide, a CTLA-4 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide, a synthetic peptide (not based on a protein associated with the immune response), or a combination thereof.
29 . The CAR of claim 27 , wherein the at least one signaling region comprises a CD3ζ polypeptide and a co-stimulatory domain, which optionally comprises a 4-1BB polypeptide or a CD28 polypeptide.
30 . The CAR of claim 27 , wherein the at least one signaling region comprises a co-stimulatory signaling region.
31 . The CAR of claim 30 , wherein the co-stimulatory signaling region comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, or a combination thereof.
32 . The CAR of claim 31 , wherein the co-stimulatory signaling region comprises a CD28 polypeptide or a 4-1BB polypeptide.
33 . The CAR of claim 32 , wherein
(a) the transmembrane domain comprises a CD28 polypeptide, the intracellular domain comprises a CD3ζ polypeptide, and the co-stimulatory signaling domain comprises a CD28 polypeptide; or
(b) the transmembrane domain comprises a CD8 polypeptide, the intracellular domain comprises a CD3ζ polypeptide, and the co-stimulatory signaling domain comprises a 4-1BB polypeptide.
34 . The CAR of claim 1 , wherein the CAR is recombinantly expressed or expressed from a vector.
35 . An immunoresponsive cell comprising the CAR of claim 1 .
36 . The immunoresponsive cell of claim 35 , wherein the CAR is constitutively expressed on the surface of the immunoresponsive cell.
37 . The immunoresponsive cell of claim 35 , wherein the isolated immunoresponsive cell is further transduced with at least one co-stimulatory ligand such that the immunoresponsive cell expresses the at least one co-stimulatory ligand.
38 . The immunoresponsive cell of claim 35 , wherein the immunoresponsive cell is selected from the group consisting of a T cell, a Natural Killer (NK) cell, a cytotoxic T lymphocyte (CTL), a regulatory T cell, a human embryonic stem cell, a lymphoid progenitor cell, a T cell-precursor cell, and a pluripotent stem cell from which lymphoid cells may be differentiated.
39 . A nucleic acid molecule encoding the chimeric antigen receptor (CAR) of claim 1 .
40 . The nucleic acid molecule of claim 39 , comprising the nucleotide sequence set forth in SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, or SEQ ID NO: 235, or a sequence that is at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% homologous thereto.
41 . A vector comprising the nucleic acid molecule of claim 39 .
42 . A host cell comprising the nucleic acid molecule of claim 39 .
43 . A method of reducing tumor burden in a subject, or of reducing or lengthening survival of a subject having a neoplasm, the method comprising administering an effective amount of the immunoresponsive cell of claim 35 to the subject.
44 . The method of claim 43 , wherein the method reduces the number of tumor cells, reduces tumor size, or eradicates the tumor in the subject.
45 . The method of claim 43 , wherein the tumor or neoplasm is selected from the group consisting of multiple myeloma, Non-Hodgkin Lymphoma, Hodgkin Lymphoma, Chronic Lymphocytic Leukemia (CLL), glioblastoma, and Waldenstrom's Macroglobulinemia.
46 . A method for producing an immunoresponsive cell that binds to B cell maturation antigen (BCMA), comprising introducing into the immunoresponsive cell a nucleic acid sequence that encodes the chimeric antigen receptor (CAR) of claim 1 .
47 . A pharmaceutical composition comprising an effective amount of the immunoresponsive cell of claim 35 and a pharmaceutically acceptable excipient.
48 . A kit for treating a neoplasm, the kit comprising the immunoresponsive cell of claim 35 and written instructions for using the immunoresponsive cell for treating a subject having a neoplasm.
49 . The kit of claim 48 , wherein the neoplasm is selected from the group consisting of multiple myeloma, Non-Hodgkin Lymphoma, Hodgkin Lymphoma, Chronic Lymphocytic Leukemia (CLL), glioblastoma, and Waldenstrom's Macroglobulinemia.
50 . The CAR of claim 1 , wherein the extracellular antigen-binding domain is a single-chain variable fragment (scFv), a Fab that is optionally crosslinked, or a F(ab) 2 .
51 . The CAR of claim 1 , wherein the extracellular antigen-binding domain is a human scFv, and/or the heavy chain variable region is a human heavy chain variable region, and/or the light chain variable region is a human light chain variable region.
52 . The CAR of claim 1 , wherein the extracellular antigen-binding domain binds to a non-linear epitope of BCMA.
53 . A CAR comprising an extracellular antigen-binding domain that specifically binds to B cell maturation antigen (BCMA), a transmembrane domain and an intracellular domain, wherein the extracellular antigen-binding domain binds to BCMA with a binding affinity (K D ) of from about 1×10 −9 M to about 3×10 −6 M and comprises a human heavy chain variable region and a human light chain variable region.
54 . The CAR of claim 53 , wherein the extracellular antigen-binding domain of the CAR binds to BCMA with a binding affinity (K D ) of from about 1×10 −9 M to about 1×10 −8 M.
55 . The CAR of claim 53 , wherein the extracellular antigen-binding domain binds to a non-linear epitope of BCMA.
56 . The CAR of claim 53 , wherein the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 21, 53, 57, 61, or 65; and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 22, 54, 58, 62, or 66.
57 . The CAR of claim 53 , wherein:
a) the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 21, and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 22;
b) the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 53 and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 54;
c) the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 57 and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 58;
d) the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 61 and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 62; or
e) the heavy chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a heavy chain variable region sequence set forth in SEQ ID NO: 65 and the light chain variable region comprises an amino acid sequence that is at least about 95% homologous to, or a CDR1, a CDR2, and a CDR3 of, a light chain variable region sequence set forth in SEQ ID NO: 66.
58 . The CAR of claim 53 , wherein:
a) the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 21 and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 22;
b) the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 53 and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 54;
c) the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 57 and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 58;
d) the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 61 and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 62; or
e) the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 65 and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 66.